OBJECTIVE: Chapter 1: Investigate the impact that trimethylamine N-oxide (TMAO), dietary contribution of short chain fatty acids (SCFAs), and role of bile acids has on cardiovascular health and disease. Chapter 2: Evaluate the association between intakes of dietary protein from both animal and plant sources on lipid profile changes. METHODS: Chapter 1: Literature review using PubMed and EMBASE to search for published studies for dietary intake or supplementation impact on TMAO or its precursors and their role in the development or prevention of cardiovascular diseases. Chapter 2: Framingham Offspring Study, prospective cohort study using statistical methods to investigate the changes in lipid profiles with dietary animal and plant protein. PUBLISHED STUDIES/RESULTS: Chapter 1: The increased risk of cardiovascular diseases (CVD) correlates with increasing levels of circulating levels of TMAO. The risk of CVD in animal and human studies have shown to be distinct in groups with and without CVD, leading to either beneficial or adverse effects from the consumption of dietary phosphatidylcholine, choline, betaine, carnitine, or intact TMAO. A Western dietary approach has been linked with the development of dyslipidemia whereas, adherence to a Mediterranean diet reduces the risk of major CVD events. The dietary precursors involved in TMA production by the gut microbiota then respectively to TMAO through hepatic enzyme FMO3 provide both beneficial and detrimental effects. Mechanisms of action for TMAO on CVD risk involves changes associated with cholesterol and sterol metabolism leading to foam cell formation, and enhancement of scavenger receptors, CD36 and scavenger receptor-A, on macrophages affects the rate of cholesterol influx and efflux. Choline derived in a dose-dependent manner from eggs improves cardiometabolic biomarkers with no changes in fasting TMAO. Further, choline from eggs also increases the lipoprotein particle size for both HDL-cholesterol and LDL-cholesterol increasing the rate of reverse cholesterol transport (RCT). Betaine concentrations in humans are associated with health outcomes based on an individual’s overall systemic health at baseline. Supplementation with L-carnitine produced favorable effects in lean subjects compared to obese subjects, improved cardiometabolic status in patients with myocardial infarction, and improved lipid profiles among individuals with prevalent coronary heart disease (CAD). Fish consumption increases concentrations of TMAO due to its high levels of intact TMAO though, protective effects for CVD are obtained from fatty fish providing omega-3-fatty acids impacting positive changes in the lipid profiles. Antibiotic therapy suppresses the gut microbiota and eliminates the production of TMA from the dietary precursors that are required. Chapter 2: Men and women both showed a decreasing trend for LDL-cholesterol as the tertiles increased for animal protein intake. Plant protein intake showed a similar decreasing trend for LDL-cholesterol with increasing protein tertiles; however, men had inconsistency among the trend whereas women had a consistent decreasing trend. HDL-cholesterol content increases in males and females with both increasing tertiles for animal and plant protein, though plant protein presented much stronger effects when compared to animal protein. Log-transformed triglycerides were inversely associated with increasing animal protein intake, men revealing greater effects than females. Plant protein intake showed a stronger effect than animal protein intake in an increasing trend in the log of triglycerides over the 6 exams. Overall, total cholesterol content varied at each examination period, animal protein intake tertiles displayed decreased level of total cholesterol, there was a greater effect in men than women. Higher intake of plant protein had a similar trend to animal protein intake showing a decrease in the total cholesterol concentration. Women had a much greater effect in reducing total cholesterol with plant protein when compared to men. CONCLUSION: Chapter 1: Multiple human and animal trials addressed in the association between diet, dietary precursors, gut microbiota composition, and their derived metabolite TMAO on the presence or absence of CVD display contradictory results and identifies areas needing further study. Chapter 2: Regardless of the source of protein, the lipid profiles improved with the intake of either animal or plant protein as the protein intake was increased over the tertiles in each exam. The overall trend with increasing animal or plant protein intake led to decrease in LDL-cholesterol, log transformed triglycerides, and total cholesterol whereas, the HDL-cholesterol concentrations were increased. Men favored animal protein intake to show greater reductions in LDL-cholesterol and total cholesterol, whereas women favored plant protein. The increase in HDL-cholesterol concentration was stronger with the intake of plant protein in men and women. The changes in log transformed triglycerides were similar in men and women.